Discovery of Terminal Oxazole-Bearing Natural Products by a Targeted Metabologenomic Approach

A targeted metabologenomic method was developed to selectively discover terminal oxazole-bearing natural products from bacteria. For this, genes encoding oxazole cyclase, a key enzyme in terminal oxazole biosynthesis, were chosen as the genomic signature to screen bacterial strains that may produce oxazole-bearing compounds. Sixteen strains were identified from the screening of a bacterial DNA library (1,000 strains) using oxazole cyclase gene-targeting polymerase chain reaction (PCR) primers. The PCR amplicon sequences were subjected to phylogenetic analysis and classified into nine clades. ¹H−¹³C coupled-HSQC NMR spectra obtained from the culture extracts of the hit strains enabled the unequivocal detection of the target compounds, including five new oxazole compounds, based on the unique ¹J_CH values and chemical shifts of oxazole: lenzioxazole ( 1 ) possessing an unprecedented cyclopentane, permafroxazole ( 2 ) bearing a tetraene conjugated with carboxylic acid, tenebriazine ( 3 ) incorporating two modified amino acids, and methyl-oxazolomycins A and B ( 4 and 5 ). Tenebriazine displayed inhibitory activity against pathogenic fungi, whereas methyl-oxazolomycins A and B ( 4 and 5 ) selectively showed anti-proliferative activity against estrogen receptor-positive breast cancer cells. This metabologenomic method enables the logical and efficient discovery of new microbial natural products with a target structural motif without the need for isotopic labeling.